Presentation on theme: "HLTH 340 Lecture A3 Toxicokinetic processes: absorption (part-2) carrier-mediated transport NOTICE: These materials are subject to Canadian copyright."— Presentation transcript:
1HLTH 340 Lecture A3 Toxicokinetic processes: absorption (part-2) carrier-mediated transport NOTICE: These materials are subject to Canadian copyright and are presented here as images published in journals and books for which the University of Waterloo holds a licensed electronic subscription. These materials are provided to HLTH 340 students for their exclusive use though a non-public courseware system (UW-LEARN) and the images are restricted to the use of HLTH 340 students. Reproduction, transmittal, copying, or posting of these images by students in any form, electronic or physical, is strictly prohibited.
2Carrier-mediated absorption using membrane transport channels large glycoprotein molecules embedded in the phospholipid membraneact as channels (pores) that allow specific hydrophilic solutes (e.g. metal ions) to cross membrane barriersenables transcellular absorption of many hydrophilic ions and polar molecules2 types of membrane transporter channelsactive transport channelsfacilitated diffusion channelsactive transportincreases rate of absorptioncan concentrate (or remove) substance in tissues against a concentration gradient of a given solute (non-equilibrium; pumps solutes ‘uphill’)requires energy source (e.g. ATP or other source of chemical energy)facilitated diffusion (passive transport)does not concentrate or eliminate substance in tissues (equilibrium only; no pumping)no energy source required
4Absorption of inorganic ions through membrane ion transport channels each type of channel has a selective preference for transporting certain ionsspecific solutes are preferred, but not absolutely specificfactors determining ion transporter specificity include• ionic radius(size)• positive or negative charge• valency of the ionic charge• class of metal ion (alkaline metals, transition metals, heavy metals)size and ionic charge determines selective transport thru ion channelscation transport channels (K+, Na+, Ca++, etc.)transport positively charged ions (e.g. metals)monovalent (+1), divalent (+2), trivalent (+3), etc.anion transport channels (Cl-, I- , ClO3- etc.)transport negatively charged ionsmonovalent (-1), divalent (-2), trivalent (-3), etc.factors affecting rate of transportaffinity (high-affinity ==> rapid transport) <-- determined mainly by ionic charge and the effective ionic radiussaturation (rate-limiting factor for absorption speed)competition (2 substances compete for the same transporter channel)regulation (up-regulation or down-regulation by other physiological factors)
5Metal speciationmetals can exist in different physico-chemical states (species)elemental (metallic)oxidized (metal oxides and salts)inorganic or organic compoundsmetal speciationinorganic metals can have several different oxidation stateselemental (metallic) species, Meoneutral, no electric chargesolid metal (e.g. Pbo) or metal vapor (e.g. Hgo )oxidation states: loss of electron (LEO) is chemical oxidationMe+ loss of 1 electron is valence state (I)Me++ loss of 2 electrons is valence state (II)Me+++ loss of 3 electrons is valence state (II)different species of the same metal can vary widely in toxicityexample: chromium: Cr-III (chromium three) -- oxidation--> Cr-VI (chromium six) low toxicity high toxicityorganometallic compoundsmetal atom covalent linked to organic (carbon chain) group(s)frequently very toxiclipophilic organometal compounds are especially hazardousexample: mercurymercuric ion (Hg2+) --> methyl mercury (organic)low toxicity high toxicity
6Metal ions - ‘heavy metal’ ions compete for membrane transport via ionic mimicry precious metals (toxic, but rare)essential metalstrivalent metals(too ionic or insoluble to be absorbed)toxic metalsheavy metals (ionic mimicry)transition metals (absorbed via transport channels)alkali metals (absorbed via transport channels)6
7Intestinal calcium transport channels: calcium (Ca++) uptake competes with lead (Pb++) intestinal epithelium contains selective ion transport channels for different divalent cations (Ca++, Fe++, Zn++)each serves as a transcellular transport channel of divalent cations from the intestinal lumen into the bloodstreamexample: calcium (Ca++ )calcium is an essential metal nutrientparacellular uptake by passive diffusion is sufficient at high dietary calcium levels(strong conc gradient)transcellular carrier-mediated transport is required for low dietary concentrations of calciumCa++ uptake requires transcellular transport via a selective divalent cation calcium channelrequires ATP energy to run the active transport system (pump)selective calcium absorption occurs from many foods (milk, dairy products, vegetables, fruits, fish)TRPV6 (ECaC2) calcium channelCa++ is absorbed from the intestine to blood using indirect active transport via the TRPV6 calcium channelepithelial calcium channel type 2 (ECaC2; now termed TRPV6)carrier-mediated uptake by TRPV6 is up-regulated by active vitamin D 1, 25-(OH)2 D3• sunlight / UV synthesis / melanization • vitamin D supplements in milk and other diary productsalso sustained by estrogen hormones (up-regulation)** **ocurs only in femaleslead (Pb++ )lead exists as a divalent cation (Pb++) -- the ion structure of lead mimics calcium ion (ionic mimicry)Pb++ is absorbed from the intestine into the blood using same Ca++ transport channel (TRPV6 )saturation-competition occurs with calcium versus lead for saturable TRPV6 channels in intestine7
8Carrier-mediated transport is a saturable process that has a limited capacity to move solutes paracellular intestinalcalcium absorptionat high Ca++ conc (passive diffusion)TRPV6 transport channel becomes saturated at high Ca++ concTRPV6 intestinalcalcium absorptionat low Ca++ conc (carrier-mediated)8
9Calcium ion transport channel embedded in epithelial cell membrane (TRPV6) Pb2+9
10Organized water - ionic groups and polar groups effective ionic radius - ionic molecules and metal ions form electrostatic bonds with the surrounding water molecules, so that the ions actually have a much larger sphere of hydration than the notional size of the unhydrated metal ion -- the radius of the sphere of hydration determines the degree of penetration of the metal ion through a specific carrier-mediated transport channelhydrogen bonding - in hydrophilic polar molecules, the hydrogen in one or more functional groups (e.g. -OH) has a slight positive charge and is therefore attracted by the partly negatively charged oxygen of a nearby water molecule, with the result that a hydrogen bond is formed.The dissolved (solute) molecules and the water molecules thus become linked to one another. When non-ionic polar molecules form hydrogen bonds with the surrounding water molecules, the polar molecules are ‘glued’ to the water solvent layer -- this organized water layer must be stripped away before the solute molecule can penetrate through a narrow carrier-mediated transport channel10
11Ion transport channels - the effective ionic radius of hydration sphere for divalent cations effective ionic radius - for ionic radius of metal divalent cations, the smaller ions have higher ionic potentials and form stronger bonds with water molecules, so that smaller ions actually have a larger sphere of hydration when entering into a specific ion transport channel11
12Structure of the TRPV6 (ECaC2) tetrameric calcium channel using molecular modeling IG. 10. Structure of the TRPV5/6 pore. A: alignment of the pore regions of TRPV4, -5, and -6 and the potassium channel KcsA (accession numbers AAG17543 for TRPV4, CAB40138 for TRPV5, AAD47636 for TRPV6, and S60172 for KcsA). The boxed sequences represent the putative selectivity of all channels. The amino-terminal part of this pore structure likely represents a putative pore helix. Band C: model for the TRPV6 pore region, based on the KcsA structure. Views of the structure are shown, looking sideways at two opposite subunits ( left ) or looking down from the external solution to the complete homotetrameric channel ( right ). At the narrowest point, formed by the acidic side chain of Asp-541 (orange), the pore has a diameter of Blue residues correspond to the residues in TM5 and TM6 (TM1 and TM2 in KcsA), whose accessibility was not tested in this study. Amino acids that were subjected to SCAM analysis (residues P526 to D547) are colored in green, yellow, red, or gray: residues in red reacted rapidly to Ag +(reaction rate >5 x10 6M–1 · s –1 ), residues in yellow reacted with Ag +at a rate <5 x10 6M–1 · s –1 , and residues in green did not show significant reactivity to Ag +. Residues where cysteine substitution resulted in nonfunctional channels are marked in gray. The figure was prepared using DS ViewerPro 5.0 (Accelrys), based on the coordinates of KcsA.12
13ATP-dependent channel Cation transport channel (TRPV6) for calcium ion: absorption in the intestinal epitheliumantiporter channelPb++ATP-dependent channelFIG. 1. Mechanism of epithelial Ca 2+ transport. Epithelia can absorb Ca 2+ by paracellular and transcellular transport. Passive and paracellular Ca 2+ transport takes place across the tight junctions and is driven by the electrochemical gradient for Ca 2+ (blue arrow). The active form of vitamin D, 1,25-dihydroxyvitamin D 3[1,25-(OH) 2D3], stimulates the individual steps of transcellular Ca 2+ transport by increasing the expression levels of the luminal Ca 2+ channels, calbindins, and the extrusion systems. Active and transcellular Ca 2+ transport is carried out as a three-step process. Following entry of Ca 2+ through the (hetero)tetrameric epithelial Ca 2+ channels, TRPV5 and TRPV6, Ca 2+ bound to calbindin diffuses to the basolateral membrane. At the basolateral membrane, Ca 2+ is extruded via a ATP-dependent Ca 2+ -ATPase (PMCA1b) and a Na +/Ca 2+ exchanger (NCX1). In this way, there is net Ca 2+ absorption from the luminal space to the extracellular compartment13
14Absorption pathways for lead permeation from the intestinal lumen through the enterocytes TRPV6(Ca++)??DMT-1 (Fe++)transcellularFIG. 1. Mechanism of epithelial Ca 2+ transport. Epithelia can absorb Ca 2+ by paracellular and transcellular transport. Passive and paracellular Ca 2+ transport takes place across the tight junctions and is driven by the electrochemical gradient for Ca 2+ (blue arrow). The active form of vitamin D, 1,25-dihydroxyvitamin D 3[1,25-(OH) 2D3], stimulates the individual steps of transcellular Ca 2+ transport by increasing the expression levels of the luminal Ca 2+ channels, calbindins, and the extrusion systems. Active and transcellular Ca 2+ transport is carried out as a three-step process. Following entry of Ca 2+ through the (hetero)tetrameric epithelial Ca 2+ channels, TRPV5 and TRPV6, Ca 2+ bound to calbindin diffuses to the basolateral membrane. At the basolateral membrane, Ca 2+ is extruded via a ATP-dependent Ca 2+ -ATPase (PMCA1b) and a Na +/Ca 2+ exchanger (NCX1). In this way, there is net Ca 2+ absorption from the luminal space to the extracellular compartmentZIP-1 (Zn++)paracellular14
15Lead (Pb): a heavy metal toxicant that is a common environmental health hazard 4 major toxic heavy metals are of common concern in environmental healthleadmercurycadmiumarsenicmajor environmental sources of leadleaded paints (lead oxide PbO) -- houses built before 1978auto exhaust in soils (PbCO3 from leaded gasoline) -- leaded gasoline additive (tetraethyl lead) until 1976pica in young children (PbO in house dust, paint chips)lead contamination dust (industrial and lead smelter neighborhoods)lead drinking water pipes in older neighborhoods/houseschronic exposure to very low levels of lead (~10 ug/dL??) can cause permanent neurologic damage in childrenIQ reduction and subtle mental retardationhyperactivity and behavioral disorders (maybe??)prevention of chronic lead poisoning -- (1) infants (2) pregnant women (3) elderlycalcium supplements (BEWARE!)diet with dairy products, high protein intake, adequate vitamin Destrogen supplements in post-menopausal women??
16Pathological features of lead poisoning in adults and children In the adult, the brain and central nervous system are relatively less vulnerable to the toxic effects of lead because the adult brain is biologically matured and does not undergo developmental retardation as is the case for infants and young children.Therefore many adults may experience other health effects such as chronic anemia, hypertension, or kidney malfunction.In children the main effect is neurological (impaired brain development).16